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1.
目的:研究NF-κB"诱骗(decoy)"寡核苷酸(ODNs)对脂多糖(LPS)诱导小鼠巨噬细胞株J774.1细胞诱导型一氧化氮合酶(iNOS)表达的影响.方法:体外培养巨噬细胞,LPS刺激后脂质体转染NF-κB"decoy"ODNs,用硝酸还原酶法测定培养液中NO水平以及iNOS含量,RT-PCR观察细胞iNOS的mRNA表达变化.结果:NF-κB"decoy"ODNs可降低巨噬细胞培养体系LPS诱导的NO合成,阻断iNOS mRNA的表达;对照序列的ODNs和转染剂对NO、iNOS无明显影响.结论:NF-κB"decoy"ODNs可以抑制iNOS的活性,减少NO生成,可能与其特异性竞争抑制活化NF-κB结合位点有关.  相似文献   

2.
博来霉素和石棉对肺泡巨噬细胞转录因子的活化作用   总被引:1,自引:1,他引:0  
目的研究博来霉素和石棉对THP-1和肺泡巨噬细胞内转录因子核因子-κB (NF-κB)和环磷酸腺苷效应元件(CREB)的活化作用,探讨肺泡巨噬细胞内转录因子在肺间质性疾病中的作用.方法用磷酯酰多糖(LPS)或石棉刺激THP-1细胞,提取细胞核蛋白质,以电泳迁移率变动分析方法检测NF-κB和CREB的活性.采用纤维支气管镜进行支气管肺泡灌洗收集健康非吸烟者的肺泡巨噬细胞,用LPS、博来霉素及石棉分别或共同刺激细胞,提取核蛋白质,用电泳迁移率变动分析方法检测NF-κB的活性.以竞争抑制实验检测DNA结合蛋白的特异性.结果 THP-1细胞未受到刺激时,转录因子NF-κB和CREB均具有一定的基础活性;在LPS或石棉刺激后3、6及24 h后,NF-κB的活性进一步增强,但CREB的活性无明显变化. 在未刺激的肺泡巨噬细胞中,存在NF-κB的基础活性,在LPS刺激后NF-κB的活性明显增强,3 h为肺泡巨噬细胞NF-κB活化增强的最佳时间,24 h时NF-κB活性下降.博来霉素(10-2 mmol/ml)或石棉(100 μg/ml)均可诱导NF-κB的活性增强,LPS可进一步增强博来霉素或石棉对NF-κB的活化作用.结论在博来霉素或石棉引起的肺间质疾病中,肺泡巨噬细胞转录因子NF-κB的活化可能具有重要的作用.  相似文献   

3.
豚鼠椎基底动脉缺血/再灌注耳蜗损伤NF-кBp65表达的研究   总被引:1,自引:0,他引:1  
刘斌  任基浩  葛益林   《中国医学工程》2008,16(2):110-112
目的探讨豚鼠椎基底动脉缺血/再灌注时是否存在NF-κBp65的激活。方法经颅底径路建立豚鼠椎基底动脉缺血/再灌注耳蜗损伤模型,采用石蜡包埋免疫组织化学技术,对各组动物近中轴位切片行NF-κBp65免疫组织化学染色,光学显微镜下观察在耳蜗组织中的表达情况;提取缺血再灌注侧耳蜗组织的细胞核和细胞浆蛋白,应用电泳迁移率滞后分析检测细胞核内NF-κBDNA结合活性。结果正常豚鼠耳蜗螺旋神经节、Cord’s器、血管纹、螺旋韧带的细胞浆和细胞核内NF-κBp65染色呈阴性,缺血再灌注12h可见明显表达,24h表达最强,48h表达逐渐减弱;正常耳蜗组织中可见少量的NF-κB活化,在缺血再灌注的耳蜗组织中NF-κBDNA的结合活性显著上调,再灌注24h的结合活性最高,再灌注48h后NF-κBDNA的结合活性下降。结论耳蜗缺血再灌注损伤时,NF-κBp65在耳蜗出现异常表达,NF-κBDNA结合活性增高。  相似文献   

4.
目的 研究替米沙坦对脂多糖诱导3T1-L1脂肪细胞炎症通路的影响,探讨替米沙坦的抗炎机制.方法将3T3-L1脂肪细胞诱导至90%成熟,按随机数字表法分为6组:对照组、LPS组、LPS+替米沙坦(0.01、0.1、1和10μg/ml)组(n=18),向对照组细胞中加DMSO,LPS+替米沙坦组细胞中加入不同浓度替米沙坦,孵育2h后,LPS组和LPS+替米沙坦组再加入脂多糖(1 μg/ml),孵育1h,Western免疫印记法比较各组核蛋白NF-κBp65、总蛋白p-IκBα、IκBα、p-ERK1/2、ERK1/2、P-p38MAPK、p38 MAPK的表达水平.结果 脂多糖刺激后IκBα磷酸化蛋白表达增强(P<0.05),细胞核内NF-κBp65蛋白表达增多(P<0.05),MAPK通路相关蛋白ERK1/2磷酸化和p38MAPK磷酸化水平明显增强(P<0.05),替米沙坦可抑制IκBα磷酸化和NF-κBp65核转位,p-IκBα蛋白表达和核蛋白NF-κBp65表达均降低,大剂量替米沙坦(10 μg/ml)作用时该作用明显(P<0.05).替米沙坦干预后,p-ERK1/2和p-p38MAPK蛋白表达也受到明显抑制(P<0.05),大剂量(10μg/ml)时作用明显(P<0.05).结论 在3T3-L1脂肪细胞中,替米沙坦可以抑制NF-κB通路中IκBα蛋白磷酸化、NF-κBp65蛋白核转位以及MAPK通路中ERK1/2蛋白和p38MAPK蛋白的磷酸化,从而发挥抗炎作用.  相似文献   

5.
目的 探讨小于扰RNA(siRNA)抑制核因子κB(NF-κB)信号通路对细菌脂多糖(LPS)刺激的巨噬细胞释放肿瘤坏死因子α(TNF-α)的影响.方法 人单核细胞白血病THP-1细胞经氟波酯诱导为巨噬细胞后分2组,分别感染应用分子克隆技术构建的针对NF-κB P65的siRNA逆转录病毒质粒(siRNA病毒组)及乱序(Scramble)对照序列逆转录病毒质粒(Scramble病毒组).用终浓度50μg/ml的LPS持续刺激2组巨噬细胞,分别应用RT-PCR技术检测LPS刺激不同时间巨噬细胞NF-κB P65 mRNA和TNF-α mRNA表达水平,蛋白质印迹法检测NF-κB P65蛋白表达水平,酶联免疫吸附试验检测巨噬细胞培养上清液TNF-α含量.结果 LPS刺激12和24 h,siRNA病毒组巨噬细胞NF-κB P65 mRNA表达水平(0.97±0.02,0.89±0.01)明显低于Scramble病毒组(1.01±0.03,0.97±0.01,均P<0.05);LPS刺激24和72 h,siRNA病毒组NF-κB P65蛋白表达水平(0.95±0.04,0.94±0.01)明显低于Scramble病毒组(1.07±0.06,1.03±0.05,均P<0.05).LPS刺激4、8、12和24 h的各时点,siRNA病毒组TNF-α mRNA表达水平均明显低于Scramble病毒组(0.92±0.02比0.98 ±-0.01,0.86±0.02比1.00±0.01,0.79±0.03比1.01±0.01,0.78±0.03比1.02±0.01,均P<0.05).LPS刺激2、4、8、24、36、48、54和72 h的各时点,siRNA病毒组巨噬细胞培养上清液TNF-α含量均明显低于Sramble病毒组(均P<0.01).结论 针对NF-κB P65的siRNA可抑制NF-κB信号通路的功能活性,进而抑制内毒素刺激引起的巨噬细胞活化和TNF-α释放,提示RNA干扰技术在急性肺损伤早期过度炎症反应的防治中具有潜在应用价值.  相似文献   

6.
目的探讨单核细胞NF-κB的激活在COPD发病机制中的作用。方法抽取31例急性加重期组和26例稳定期组的COPD患者外周血,分离单核细胞并培养,免疫细胞化学法测定核转录因子NF-κBp65的活性表达;同时检测外周血SOD、MDA、IL-8浓度,所有患者均测定肺功能(FEV1占预计值%、FEV1/FVC)。并以15例健康志愿者作为对照组。结果LPS刺激后,急性加重期组单核细胞NF-κBp65表达显著高于稳定期组及正常组(P<0.01),稳定期组高于正常组(P<0.01)。经LPS刺激后,各组单个核细胞NF-κBp65活性均显著增高,(P<0.05);外周血单核细胞NF-κB的激活与血清IL-8、MDA水平呈正相关,而与SOD呈负相关。结论NF-κB在COPD单核细胞表达细胞因子IL-8的信号转导调控中起重要作用。  相似文献   

7.
目的 研究内毒素(lipopolysaccharide,LPS)尾静脉注射诱发ALI大鼠动物模型中肺组织核因子-κBp65(nuclear factor-kappaB 65,NF-κBp65)的表达,用清开灵干预,观察和探讨清开灵对急性肺损伤(acute lung injury,ALI)的治疗效果及作用机制.方法 30只大鼠,随机分成5组,每组6只.对照组(A组):706代血浆每只2 mL/kg尾静脉注射;模型组(B组):LPS每只6 mg/kg溶于706代血浆,每只2mL/kg尾静脉注射;低剂量清开灵组(C组):LPS用法同B组,2 h后尾静脉每只注射清开灵2 mL/kg;中剂量清开灵组(D组):LPS用法同B组,2 h后尾静脉注射清开灵每只3 mL/kg;高剂量清开灵组(E组):LPS用法同B组,2 h后尾静脉注射清开灵每只4 mL/kg.观察各组动物反应,清开灵干预2 h后颈动脉快速放血处死,行动脉血气分析,剖胸取右肺称湿重后,90℃烘箱内24h至恒重,称干重,计算湿/干重比(wet weight/dry weight,W/D).左肺用4%中性甲醛溶液固定、石蜡包埋、组织切片(5μm),HE染色,行病理观察,免疫组化及图像分析系统检测NF-κB65的表达.结果 各清开灵治疗组能不同程度减轻呼吸窘迫,模型组肺组织呈暗红色,肺组织增大,明显充血,水肿,可见广泛的点状或灶状出血;清开灵干预后,充血,水肿减轻.光镜下模型组可见肺灶性出血,肺泡壁增厚,肺泡和肺间质水肿,大量炎性细胞浸润,各剂量清开灵干预组病理学改变较模型组轻.结论 ALI大鼠肺组织中NF-κBp65表达明显升高,ALI的发病机理可能与NF-κBp65表达升高所致的炎症反应调控失衡有关;清开灵对大鼠ALI有明显的治疗效果,可能与其下调NF-κBp65表达有关.  相似文献   

8.
蓝桉油对单核细胞THP-1核因子-κB核转位活化的影响   总被引:2,自引:0,他引:2  
目的:探讨蓝桉油对人单核细胞THP-1细胞株核因子-κB(NF-κB)核转位活化的影响。方法:蓝桉油(1、10、100mg/L,30min)预处理THP—1细胞,经脂多糖(LPS,1mg/L,30min)刺激后,采用间接免疫荧光细胞化学法结合激光扫描共聚焦显微镜,测定THP—1细胞NF-κBp65亚基(NF-κB/p65)定位;Western-blot法分析细胞核内NF-κB/p65水平。结果:免疫荧光分析显示,正常细胞NF-κB/p65荧光标记主要分布于胞浆区,核区很少,LPS刺激后NF-κB/p65荧光标记浓集于核区,胞浆区少见;但经蓝桉油(100mg/L)预处理后,LPS刺激的THP—1细胞NF-κB/p65荧光标记则主要位于胞浆区;Western-blot分析显示,在正常THP—1细胞核蛋白中有低水平的NF-κB/p65表达,经LPS刺激,核内NF-κB/p65表达明显增高;蓝桉油预处理后,可以抑制LPS诱导的核内NF-κB/p65高水平表达,且呈剂量依赖关系。结论:蓝桉油预处理阻止了LPS诱导的NF-κB/p65核转位,从而抑制了其活性功能的发挥。蓝桉油可抑制LPS刺激引起的NF-κB核转位活化。  相似文献   

9.
目的研究肺泡巨噬细胞在脂多糖(LPS)作用条件下一氧化氮供体(SNP)对糖皮质激素受体(GR)功能的调节作用.方法将GR荧光表达质粒pGFP-GR转染大鼠肺泡巨噬细胞,经LPS和SNP作用后,荧光显微镜观察质粒表达产物GFP-GR核移位情况;相对荧光素酶法检测GR转录激活活性;EMSA检测检测细胞NF-κB活性.结果 500 μmol/L SNP作用2 h出现GFP-GR核移位,同时GR转录激活活性显著增强,NF-κB活性被显著抑制.运用GR特异性拮抗剂RU486后,NF-κB活性抑制现象消失.结论肺泡巨噬细胞在致炎因子作用条件下,一氧化氮供体(500 μmol/L SNP )通过活化GR发挥抗炎活性.  相似文献   

10.
目的 研究核因子κB p65(NF-кB p65)对氧化低密度脂蛋白(OX-LDL)处理的内皮祖细胞增殖活性的影响及机制。方法 分离培养人内皮祖细胞,用OX-LDL细胞培养液培养后,分别用实时荧光聚合酶链反应(qRT-PCR)和Western blotting检测细胞中NF-κBp65 mRNA及蛋白水平。用NF-κB p65 siRNA慢病毒感染内皮祖细胞,给予OX-LDL处理以后,qRT-PCR和Western blotting检测干扰效果。MTT检测内皮祖细胞的增殖活性,流式细胞术检测内皮祖细胞的凋亡变化,Western blotting检测细胞中活化的Caspase-3、Caspase-9蛋白水平,用比色法检测细胞裂解液中的超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量。结果 OX-LDL处理可以诱导内皮祖细胞中NF-κB p65表达上调。NF-κB p65 siRNA慢病毒可以下调OX-LDL条件下内皮祖细胞中NF-кBp65的表达水平。OX-LDL处理后的内皮祖细胞的增殖活性降低,细胞凋亡率升高,细胞中活化的Caspase-3、Caspase-9蛋白水平也升高,细胞中SOD活性降低,MDA含量升高(P?<0.05)。敲低NF-κB p65 后的内皮祖细胞经OX-LDL诱导后,细胞增殖活性有所升高,细胞凋亡率降低,细胞中活化的Caspase-3、Caspase-9蛋白水平降低,SOD活性升高,MDA含量降低(P?<0.05)。结论 敲低NF-κB p65能够降低OX-LDL诱导的氧化应激及细胞凋亡,提高内皮祖细胞增殖活性。  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

14.
目的:评价使用安心颗粒对急诊经皮冠状动脉介入术(PPCI)术后生活质量的影响.方法:将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组(术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次)和对照组(仅接受基础药物治疗).所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表(MIDAS)、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果:入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异(P>0.05);出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低(P<0.05);组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低(P<0.05).两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血(P>0.05).两组发生轻度血小板减少症的患者数比较无差异(P>0.05).结论:PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.  相似文献   

15.
Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P<0.01);the MAP decreased(P<0.01)and the HR increased drastically(P<0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.  相似文献   

16.
Objective:To investigate the gene expression of osteoprotegerin(OPG) and osteoclast differentiation factor(ODF) in the bone tissue of patients with hip fracture due to osteoporosis. Methods:OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers(over 50 years old) with hip fracture(Observer Group) and 30 cases of hip facture sufferers with no osteoporosis(Control group) were analyzed with the Semi-Quantitative RT-PCR method. Results:The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion:Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.  相似文献   

17.
Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.  相似文献   

18.
Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.  相似文献   

19.
Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.  相似文献   

20.
A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care(~'4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.  相似文献   

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